Strictly Conserved Residues in Euphorbia tirucalli β‐Amyrin Cyclase: Trp612 Stabilizes Transient Cation through Cation–π Interaction and CH–π Interaction of Tyr736 with Leu734 Confers Robust Local Protein Architecture

The functions of Trp612, Leu734, and Tyr736 of Euphorbia tirucalli β‐amyrin synthase were examined. The aliphatic variants (Ala, Val, Met) of Trp612 showed almost no activity, but the aromatic variants exhibited high activities: 12.5 % of the wild‐type activity for the W612H variant, 43 % for W612F, and 63 % for W612Y. That is, the enzymatic activities of the variants increased in proportion to the increase in π‐electron density. Thus, the major function of Trp612 is to stabilize transient cations through a cation–π interaction. The Phe and Tyr variants caused a distorted folding conformation, especially at the E‐ring site, which generated the aberrantly cyclized products germanicol and lupeol. The L734G and L734A variants exhibited significantly decreased activities but yielded taraxerol in a high production ratio. The Val, Ile, and Met variants showed markedly high activities (56–78 % of wild‐type activity); therefore, appropriate steric bulk is required at this position. The aliphatic variants of Tyr736 showed markedly decreased activities, but the Phe mutant exhibited high activity (67 %), which indicates that the π electrons are critical for catalysis. Homology modeling indicated that Tyr736 and Leu734 are perpendicular to the substrate and are situated face to face, which suggests that a CH–π interaction occurs between Tyr736 and Leu734, reinforcing the protein architecture, and that Tyr736 cannot stabilize cationic intermediates through a cation–π interaction.     

Effect of rolling deformation on the structures and properties of multi‐walled carbon nanotube filled isotactic polypropylene

Isotactic polypropylene filled with various contents of multi‐walled carbon nanotubes (MWCNTs) were fabricated by the injection molding technique and then rolled at room temperature. The unrolled samples (URS) and rolled samples (RS) were characterized by X‐ray diffraction studies, scanning electron microscopy, mechanical and micromechanical tests and differential thermal analyses. Although the URS exhibit the lamellar α‐crystal with a*‐axis orientation, the RS show the same crystals with both a*‐ and c‐axis orientation, which is explained by interlamellar and intralamellar slips and lamellar destruction. Scanning electron micrographs display distinct surface morphological features for both URS and RS. While the tensile strength of RS is higher than that of URS, the Young's modulus (Y) is found to be lower than that of URS. Anisotropy in microharness (H) parallel and perpendicular to the rolled direction has been detected, although H for both samples increases with increasing MWCNT contents. The average relationship H/Y ≈ 0.18 as estimated for URS is closer to the predicted value of 0.10 for polymers than the H/Y ≈ 0.22 obtained for RS. The lamellar thickness for URS increases with increase of MWCNT content and that for RS decreases, as evaluated from both differential thermal analyses and X‐ray diffraction data. Copyright © 2011 Society of Chemical Industry     

n-Hexyl Laurate and Fourteen Related Fatty Acid Esters: New Secretory Compounds from the Julid Millipede, Anaulaciulus sp.

A total of fifteen saturated fatty acid esters were newly identified from the secretions of an unidentified Anaulaciulus sp. (Julida: Julidae). The fatty acid components of the esters were composed of normal chain acids (from C₁₀ to C₁₄) and of branched chain acids (from iso-C₁₂ to iso-C₁₅ and anteiso-C₁₅). The alcohol moieties were all composed of normal chain alcohols varying from n-butanol to n-octanol. The most abundant component found in the total esters was n-hexyl laurate (64.7%). Novel compounds identified from the millipede secretion extracts include six branched iso- and anteiso-fatty esters, an odd-numbered C₁₁-fatty acid ester, a C₁₃-fatty acid ester, and a C₇-alcohol ester, all of which were previously undescribed natural products. In addition, a characteristic mixture of benzoquinones, such as 2-methyl-1,4-benzoquinone, 2-methoxy-3-methyl-1,4-benzoquinone, 2,3-dimethoxy-1,4-benzoquinone, 2-methoxy-6-methyl-1,4-benzoquinone, and 2,3-dimethoxy-5-methyl-1,4-benzoquinone were identified from the secretions, together with trace amounts of 1,4-benzoquinone.     

Synthesis and chiroptical properties of helical poly(phenylacetylene) bearing optically active chiral oxazoline Pendants

The novel optically active (S)-4-benzyl-2-(ethynylphenyl)-oxazoline (BnEPhOx) was successfully prepared and polymerized using rhodium catalyst ([Rh(nbd)Cl]₂) to obtain the moderate molecular weight poly(phenylacetylene)s bearing chiral oxazoline derivatives with high yields (≥90%). The ¹H NMR spectra demonstrated that the resulting polymers had high stereoregular structures. Moreover, the poly(phenylacetylene)s bearing chiral oxazoline exhibited better thermal stability than poly(phenylacetylene). The resulting polymers showed higher absolute values of optical specific rotation than the monomer. The polymers also exhibited intense CD signal in the region of the π-π1 band of the conjugated polyacetylene backbone in chloroform solution. The results of specific rotation and CD spectroscopy indicated that all the polymers adopted higher-order structure with predominantly one-handed screw sense.In addition, the resulting polymers emitted fluorescence under UV irradiation.     

Euphorbia tirucalli β-Amyrin Synthase: Critical Roles of Steric Sizes at Val483 and Met729 and the CH–π Interaction between Val483 and Trp534 for Catalytic Action

The functions of Val483, Trp534, and Met729 in Euphorbia tirucalli β-amyrin synthase were revealed by comparing the enzyme activities of site-directed mutants against that of the wild type. The Gly and Ala variants with a smaller bulk size at position 483 predominantly afforded monocyclic camelliol C, which suggested that the orientation of the (3S)-2,3-oxidosqualene substrate was not appropriately arranged in the reaction cavity as a result of the decreased bulk size, leading to failure of its normal folding into the chair–chair–chair–boat–boat conformation. The Ile variant, with a somewhat larger bulk, afforded β-amyrin as the dominant product. Intriguingly, various variants of Trp534 exhibited significantly decreased enzymatic activities and provided no aberrantly cyclized products, although the aromatic Phe and Tyr residues were incorporated and the steric sizes of the aliphatic residues were altered. Therefore, the Trp534 residue does not stabilize the transient cation through a cation–π interaction. Furthermore, the Trp residue, with the largest steric bulk among all natural amino acids, is essential for high enzymatic activity. Robust CH–π complexation between the Val483 and Trp534 residues is proposed herein. Altering the steric bulk at the Met729 position afforded the pentacyclic skeletons. Thus, Met729 is positioned at the E-ring formation site. More detailed insights into the functions of the Val483, Trp534, and Met729 residues are provided by homology modeling.     

Composition Regulation by Flow Copolymerization of Methyl Methacrylate and Glycidyl Methacrylate with Free Radical Method

It is important to match the feeding ratio of comonomers to the composition ratio in the resulting copolymers as closely as possible in industrial production, where the goal is often to produce more a homogeneous composition in copolymers. In this study, a flow copolymerization system with a conventionally initiated free radical method, together with randomly selected polymerization conditions is investigated. It is succeeded in achieving a closer match between the composition ratio and feeding ratio than previously reported in the copolymerization of styrene with methyl methacrylate and of glycidyl methacrylate with methyl methacrylate, which will widen the range of applications, by precisely controlling the mixing and heating in a flow polymerization apparatus. This is confirmed by the fact that the estimated values of reactivity ratios, r₁ and r₂, which are used in the reaction kinetics of copolymerization, are close to 1.     

Asymmetric anionic polymerization of n-diphenyl-methylitaconimide with chiral ligand-organometal complex

Summary Asymmetric anionic polymerizations of achiral N-diphenylmethylitaconimide (DPII) were performed with chiral ligand — organometal complexes in toluene. The obtained poly(DPII)s exhibited molecular weights of 1.5 × 10³ to 6.3 × 10³ and specific rotation between +7.5 to −18.4° (THF). The chirality of polymer was significantly affected by the organometals and chiral ligands. The poly(DPII) obtained with (4S)-2,2′-(1-ethylpropylidene)bis(4-benzyl-4,5-dihydrooxazole) (Bnbox) / n-butyllithium (n-BuLi) exhibited the highest specific rotation (−18.4°). The optical activities of the poly(DPII)s were attributed to configurational chirality of the main chain more than conformational chirality, judged from NMR, circular dichroism (CD) and GPC analyses using UV and polarimetric detectors. Received: 10 January 2002/ Revised: 12 April 2002/ Accepted: 22 April 2002     

Intramolecular triplet energy migration in poly(2-vinylnaphthalene) and poly(naphthylalkyl methacrylate)s in solid solution: Effects of side chain length

Summary The effect of side chain length on intramolecular triplet energy migration of naphthalene containing polymers was investigated by a delayed fluorescence (DF) spectroscopy for the solid solution of the polymers. The degree of triplet energy migration depends strongly on whether the chromophores are directly attached to the main chain or not.